Process for preparation of 16alpha, 17alpha, 21-trihydroxy steroids



United States Patent of Virginia No Drawing. Filed Jan. 14, B63, Ser. No. 251,970

g 9 Claims. (Q3. 26ii--397.45)

This application is a continuation-in-part of my application Serial No. 84,989, filed January 26, 1961, and now abandoned.

This invention relates to, and has for its object the provision of, a method of preparing physiologically active steroids and more particularly to preparing steroids of the l6oc,170c,2l-lllli1y lIOXy series.

In my Patent No. 3,048,581, granted August 7, 1962, I describe various 16,17-cyclic ketal and acetal derivatives of 15a,l7oz,21tlli1ydIO-Xy steroids of the pregnane series. In addition to these ll-oxygenated derivatives, other 16,17-cyclic ketal and acetal derivatives of 1604,17, ZI-trihydroxy steroids of the pregnane series, unsubstituted in the C-ring, are also known. it is often necessary to remove the ketal or acetal group in such steroids in order to prepare the free 16,17-dihydroxy steroid for use as such or for conversion into different ketal, acetal or other cyclic derivatives. Unfortunately, it has been found that normal procedures, such as refluxing with a dilute mineral acid, are inefiective to achieve the desired hydrolysis. The instant invention, therefore, relates to a process for hydrolyzing such cyclic ketals and acetals to the free 16,17,21-trihydroxy derivative.

It has been found that concentrated formic acid is unique amongst all acids tested in effecting the desired hydrolysis of a 16,17-cyclic ketal or acetal derivative of a 16,17,2l-trihydroxy steroid (or a 2l-ester thereof) to its corresponding free 16,17,21-trihydroxy derivative in high yield. Other acids, such as hydrochloric acid, are inoperative and fail to give the desired product, whereas dilute formic acid only partially hydrolyzes the starting material.

The process of the present invention, therefore, cornprises treating a 16,17-cyclic ketal or acetal of a l6a,l7o, 2l trlhydroxy steroid of the pregnane series (or a 21- ester thereof) with concentrated formic acid. The cleavage of the acetal or ketal group is followed by forrnylation in the 16 position (and other positions containing an active free hydroxyl group, such as a Zl-hydroxyl group). The resulting formates can then hehydrolyzed in the normal manner under mild conditions to yield the desired free hydroxy steroid product.

Although the process of this invention is applicable to any 16,17-cyclic ketal or acetal derivative of a 160:,17oc,

, r'2l-trihydroxy steroid of the pregnane series (including 2-esters thereof), particularly preferred are steroids of the general formula "ice wherein the 1,2 and 6,7-positions are saturated or doublebonded; R is hydrogen, R is hydrogen, hydroxy or a-acyloxy, or together R and R is keto; each X is hydrogen, halogen, hydroxy, lower alkoXy or lower alkyl, at least one X being hydrogen; Y is hydrogen or methyl; Y is hydrogen, halogen, or methyl in either the alpha or beta position; hydroxy or acyloxy; P is lower alkyl, halo lower alkyl, carboxy lower alkyl (or a salt or ester theerof), monocyclic cycloalkyl, monocyclic aryl, monooyclic aryl lower alkyl, monocyclic heterocyclic or monocyclic heterocyclic lower ialkyl; Q is hydrogen or one of the radicals defined by P; or together with the carbon to which they are joined P and Q are cycloalkyl'or monocyelic heterocyclic.

Among the suitable starting steroids utilizable in the process of this invention may be mentioned the 16,17- cyclic acetals and ketals of:

1 6 athydroxyhydrocortisone,

loc-hydroxycortisone,

16a-hydroxyprednisolone,

16a-hydroxyprednisone,

9oc-l1210- l 6whydroxyhydrocortisones (i.e.,

9a-fi uoro-l6a-hydroxyhydrocortisone,

9a-chloro-loa-hydroxyhydrocortisone,

9a-bromo-l6a-hydroxyhydrocortisone and 9Ct-lOdO" l 6a-hydr0xyl1ydrocortisone) 9a-halo-la-hydroxycortisones,

9a-halo-l6a-hydroxyprednisolone (e.g.,

triamcinolone) 9a-halo-l6oi-hydroxyprednisones,

lZu-halol 6m-hydroxyhydrocortisones e.g.,

IZm-fiuorol 6a-hydroxyhydrocortisone) 12 a-halo- 1 6 OL-hYdI'OXYCOItlSOHES (e.g.,

12a-chloro-looi-hydroxycortisone),

IZa-halo-l6oc-hydroxyprednisolones (e.g.,

IZm-fluorol 6m-hydroxyprednioslone) lZa-halo-loa-hydroxyprednisones,

6 a-methyl- 1 6a-hydroxyhydrocortisone,

6a-methyll 6 a-hydroxycortisone,

oa-methyll a-hydroxyprednisolone,

6a-methyl-l6a-hydroxyprednisone,

2ot-methyl- 1 6oz-hydroxycortisone,

2-rnetl1yll 6a-hydroxyprednisolone,

Z-mcthyllou-hydroxyprednisone,

2a,6a-dirnethy1-1ou-hydroxyhydrocortisone,

2a,6a-dimethyl-16a-hydroxycortisone,

9u-haio-2-rnethyl-loa-hydroxy prednisolones (e.g.,

Z-methyltriamcinolone) 9a-l1alo-6 a-methyll a-hydroxyhydrocortisones (e.g.,

9a-fluoro-6a-methyl- 1 6a-hydroxyhydrocortisone) 9a-halo-6a-metyl-l6a-hydroxy-prednisolones (e.g.,

6a-methyl triamcinolone) 16a-hydroXy-6-dehydrocortisone,

16oc-hydroxy-6-dehydrohydrocortisone,

9e-halo-l6 r-hydroxy-6-dehydroprednisolones,

16a-hydroXy-9o-(lower alkyl)hydrocortisones (e.g.,

16a-hydroxy-9a-methylhydrocortisone) 16a-hydroXy-9a-(iower alkyl) cortisones (e.g.,

16a-hydroxy-9a-methylcortisone) 16a-hydroxy-9 oc- (lower alkyl) prednisolones (e. g.,

16ot-hydroxy-9 a-methylprednisolone) l6a-hydroxy-9a- (lower alkyl) prednisones,

9a-(lower alkyl) -A -pregnene-1 1a,l6ot,17oc,21-t6tr0l-3,20-

diones (e.g.,

9:1- (lower alkyl) A -pregnadiene-1 1 05,1 6u,17a,21t612l'0l- 3,20-diones, and Ila-esters thereof, particularly esters with hydrocarbon carboxylic acids having less than ten carbon atoms, 6-halo-l6a-hydroxyhydrocortisones (e.g., 6u-fiuoro-l6a-hydroxyhydrocortisone, 6,8-fiuoro-16a-hydroxyhydrocortisone and 6 a-chlorol 6 a-hydroxyhydro cortisone) 6m-ch1oro-1 6ct-hydroxyprednisolone) 6-halo-1 6a-hydroxycortisone s, 6-halo-1ou-hydroxyprednisones, 6,9a-dihalo-16a-hydroxyl1ydrocortisones (e.g., 6a,9a-difiuoro 16oc-hydroxyhydrocortisone, 6[i,9a-diflu0ro-l6a-hydroxyhydrocortisone, 6u-chloro-9 rX-fillOIO-l6OL-hYdI'OXYhYdI'OCOItlSOI16 and 65,9d-dichloro-16tx-hydroxyhydrocortisone) 6,9a-diha1o-lfiot-hydroxyoortisones, V 6,9 -dihalo-lfia-hydroxyprednisolones (e.g., 6u-iluorotriamcinolone, 6fi-fluorotriamcinolone, a-chlorotriamcinolone,

6,8-chlorotriamcinolone,

' 6-halo-9ob- (lower alkyl)-l6ot-hydroxyprednisones,

1 6a, 17 a,21-trihydroxypro gesterone,

16oz, 17a,2 l-trihydroxy- 1 -dehydroprogesterone,

16a, 17 05,2 1-trihydroxy-o-dehydroprogesterone, 6-halo-16a,110:,21-trihydroxyprogesterones (e.g.,

6 Ot-fillOTO-160C, 17 :,2 I-trihydroxyprogesterone and 6a-cbloro-16a,17u,2l-trihydroxyprogesterone) 6 halo-16a,17a,21-trihydroxy-l-dehydroprogesterone's,

6-ha1o-16a,170:,21-trihydroxy-6-dehydroprogesterones,

and 2l-esters of any of the above compounds, preferably-esters WithhYCllOCfiIbOIl carboxylic acids having less than ten carbon atoms, as exemplified by a lowerv alkanoic acid ('e.g., acetic, propionic and tert-pentanoic acid), a 'monocyclic aryl carboxylic acid (e.g., benzoic. and toluicacid), a monocyclic aryl lower alkanoic acid (e.g.,

carboxylic acid.

Among the suitable aldehydes and ketones usedin the 7 preparation of the starting materials of this invention may be mentioned lower alk-anals, such as acetaldehyde,

propanal and-hexanal; di(lower aIkyDketones, such as acetone, diethylketone, dibutylketone, niethylethylketone, and methylisobutylketone; cycloalkanones, such as cyclobutanone, cyclop'entanone, cyclohexanone, suberone, and

monocyclic carbocyclic'aromatic aldehydes, such as benzaldehyde, halobenzaldehydes .(e.g., p-chlorobenzaldehyde and p-fiuorobenzaldehyde), lower a koxybenzaldehydes (e.g.,. o-anisaldehyde), di(lower allroxy)benzaldehydes (cg, veratraldehyde), hydroxybenzaldehydes (e.g., salicylaldehydes), dihydroxybenzaldehydes (e.g., resorcyalde hyde), lower alkyl benzaldehyde -(e.g., rn-tolualdehyde and p-ethylbenzaldehyde), di(lower alkyl)benza ldehydes (e.g., o,p-dimethylbenzaldehyde), acyl-amidobenzaldehydes (e.g.,- N-acetylanthranilaldehyde), and cyanobenzaldchydes; monocyclic carbocyclic' aromatic lower alkanals, such as phenylacetaldehyde,

ct-phenylpropionaldehyde, fi-phenylpropionaldehyde, y-

phenylbutyraldehyde, and -aromatically substituted halo,

lower alkoxy, hydroxy, lower alkyl, nitro, acylamido and cyano derivatives thereofi monocyclic heterocyclic aldehydes, such'as picolinaldehydes, furfural, thiophene carbonals, and haloylower alkoxy, hydroxy; lower ,alkyl,

nitro, andcyano derivatives thereof; inonocyclic heterocyclic lower alkanals; monocyclic'carbocyclic aromatic ketones, such .as acetophenone, a,ot,a-trifluoroacetophenone, propiophenone, butyrophenone, valerophenone, isocaprophenone, halophenyl lower alkyl ltetones (e.g., p chloroacetophenone and p-chloropropiophenone), (lower alkoxy)phenyl lower alkyl ketones (e.g., p-anisylmethyl ketone), di(lower alkoxy)phenyl lower alkyl ketones, hydroxyphenyl lower .alkyl ketones, dihydroxyphenyl lower alkyl ketones -(e.g., resacetophenone), (lower alkyDphenyl lower! alkyl ketones (e.g.,,. methyl p-tolyl ketone), di(lower alkyD-phenyl lower alkyl ketones (o,p-

Xylyl methyl ketone),"nitrophenyl lower alkyl ketones (e.g., p-nitrcacetophenone), acylaniidophenyl lower alkyl ketones (e.g.,- acetyl :anilines), and cyanophenyl lower alkyl ketones; benzophenone, and mono or-bis substituted halo, lower alkoXy,'hydroXy, lower alkyl, nitro, acylamido and cyano derivatives thereof; monocyclic carbocyclic aromaticlower alkanones,- suclras 1-phenyl-3-butanone and l-pheriyl-l-pentanone, and aroniatically substituted derivatives thereof; monocyclic heterocyclic ketone-s, such as Z-acetylfuran, Z-benzoylfuran, Z-acetyl-thiophene and alloXane; monocyclic heterocyclic lower alkanones; and

oxolower alkanoic acidssuch as glyoxylic, 'pyruvic, aceto acetic, fi-ketopropionic, a-ketobutyric, levulinic, S-ketocaproic andfi-ketocaprylic acid [aswell as salts and esters thereof, such as the lower alkyl esters (egymethyl and .ethyl)]. l i

- In accordance with the process of thisinvention, such' acetals and ketals are treated with concentrated'formic.

acid, By concentrated formic acid is meant aqueous .formic acid, that is, an-aqueous solution of formic acid containing more than about formic acid, optimally greater than formic acid. The process results in phenacetic and B-phenylpropionic acid), a lower alkenoic acid,'a cycloalkane carboxylic acid and a cycloalkenecyclodexanone; mono and dicycloalkyl ketones, such as cyclohexylmethylketone and dicyclopropylketone, halohyde'herniacetal, and heptafiuorobutanal ethyl herniacetal;

. halo-lower alkanones, such as. 1,l,1.-trifluoroacetone;-

lower alkanals, suchas chloral hydrate, trifiuoroacetaldenot only the cleavage of the 'acetal .or ketal group, but also in esterificationof the resulting free 16u-hydroxy group to a formate derivative. Any other labile hydroxy group,

such as a free 2l+hydroxy group, presentin the steroid molecule is also convertedto a tomato derivative.- In

. order to increase the rateof reaction, the process is preferably conducted at an elevated temperature, optimally a temperature in the range of about.40 C. to about The formic acid ester initially. formed may then be 7 saponified by treatment'witha weak base, suchas a dilute alkali (e.g., dilute sodium hydroxide and dilute potassium.

hydroxide) or a salt of a strong basev and a weak acid (e.g.-, an alkali metal carbonate or bicarbonate, such as potassium carbonate), the reaction preferably being car-' ried out'in'an inert-atmosphere, such asunder nitrogen.

This saponification step :will. not only convert any for- .mate groups to free hydroxy derivatives .but-in addition, of course, will hydrolyze 'any other ester groupings present, such as ill-esters to theirjcorrespondingfree hy-. droxyl (eig.-, Z'I-hYdI'OXY) derivatives. The foliowing exarnples illustrate the process of this invention .(all temperatures being in" degrees centigrade):

nitrobenzaldehydes,

EXAMPLE 1 T riamcinolone from triamcinolone 16,17-acetonide 21-,acetate A solution of 200 mg. of triamcinolone 16,17-acetonide ZI-acetate in 6 ml. of 90% formic acid is allowed to remain at 42 for sixteen hours. The solvent is then removed in vacuo to furnish a residue of triamcinolone 16,21-diformate. The residue is dissolved in 20 ml. of methanol and hydrolyzed under nitrogen with 4 ml. of a aqueous oxygen-free potassium carbonate solution for 30 minutes The mixture is neutralized with 0.4 ml. of glacial acetic acid, diluted with water and concentrated in vacuo until crystals appear. Filtration furnishes about 110 mg. of pure triamcinolone, identical with an authentic sample.

EXAMPLE 2 A solution of 200 mg. of triamcinolone 16,17-acetonide in 6 ml. of 88% formic acid is placed in an oven maintained at 40-42 for a period of seventeen hours. The mixture is then concentrated to dryness in vacuo to give a residue of triamcinolone 16,21-diformate. The residue is dissolved in 20 ml. of methanol and hydrolyzed under nitrogen with 4 ml. of 10% oxygen-free'aqueous potas- EXAMPLE 7 6a-flu0r0-16a-hydr0xyprednisolone from 6oc-fluOr0-16a hydroxy-prednisolone 16,17-acet0nide 21 -acetate Following the procedure of Example 1, but substituting 200 mg. of oa-fiuoro-la-hydroxyprednisolone 16,17- acetonide 21-acetate for the triameinolone 16,17-acet- .onide ZI-acetate, Goa-fluoro-loot-hydroxyprednisolone is obtained.

siumcarbonate solution for minutes at room tempera- EXAMPLE 3 6o: fluoro 160a,] 7a,21 trihydroxyprogesterone from 60: fluoro 16u,17o ,21 trihydroxy progesterone 16,17- acetonide Following the procedure of Example 1, but substituting 200 mg. of 6a-fluoro-16u,17a,2l-trihydroxyprogesterone 16,17-acetonide for the triamcinolone 16,17-acetonide 21-acetate, 6a fiuoro-16u,17a,2l-trihydroxyprogesterone is obtained.

EXAMPLE 4 60,9u difluoro 16a hydroxyhya'rocortisone from 60,

90: difluoro 16a hydroxyhydrocortisone 16,17- acetonide 21 -acetate.

Following the procedure of Example 1 but substituting 200 mg. of 6a,9u-ditluoro-16a-hydroxyhydrocortisone 16,17-acetonide 21-acetate for the triamcinolone, 16,17- acetonide ill-acetate, 6a,9u-difluoro-loa-hydroxyhydrocortisone is obtained.

EXAMPLE 5 601,90: difluoro 160thydroxyp rednisoione from 6u,9ot-

dz'fluoro 16a hydroxyprednisolone 16,1 acetonide 21-acetate Y Following the procedure of Example 1, but substituting 200 mg. of 6a,9oz-difluor o-16u-hydroxyprednisolone 16,17-acetonide 21-acetate for the triamcinolone 16,17- acetonide 21-acetate, 6m,9ot-difluoro-lda-hydroxyprednisolone is obtained.

, EXAMPLE 6 Triamcz'nolone from triamcinolone 16,17-acet0phenonide Following the procedure of Examplel, but substitutingobtained.

EXAMPLE 10 hydroxyprednisolone 16,17-acet0nide 2 1 -acetate Following the procedure, ofExample 1, but substituting 200 mg. of 12a-fiuoro-loa-hydroxyprednisolone 16,17-acetonide 21-acetate for the triamcinolone 16,17- acetonide Zl-acetate, 12a-fiu0l0-I6OL-hYdI'OXYPI'6dTliSO1OH6 is obtained.

EXAMPLE 11 6ot-fluorotriamcinolone from 6 a-fluorotriamcinolone 16,17-acet0nide 21-acetate A solution of 1.5 g. of 6a-fiuorotriamcinolone 16,17- acetonide 21-ac etate in 45 ml. of formic acid is allowed to stand in an oven heated at 42 for sixteen hours. After removal of the solvents in vacuo the dry residue of 6a-fiuorotriamcinolone 16,21-diformate is taken up in 150 ml. of methanol and treated with stirring under nitrogen with a solution of 30 ml. of 10% oxygenfree potassium carbonate in water. After 30 minutes the mixture is neutralized with 3 ml. of glacial acetic acid, diluted with water and the methanol removed in vacuo. Crystallization ensues and the resulting crystalline pre cipitate is removed by filtration. Recrystallization of the dried crystals from methanol furnishes pure ou-fluorotriamcinolone, M.P. about 269--270 (dec.), identical with an authentic sample by infrared comparison.

The following two examples illustrate the diflferences in result obtained with concentrated and dilute formic acid, respectively, and the superiority of the former.

EXAMPLE 12 A solution of 357 mg. of triamcinolone acetonide in 10 ml. of 60% formic acid (prepared by diluting 68 ml. of 88% formic acid to ml.) is heated on the steam cone at 100 for 30 minutes. A very small amount of undissolved material is filtered amounting to less than 1 mg; To the warm filtrate, water is added to turbidity and the crystals (long needles and fine rosettes) are filtered off after one-half hour in the refrigerator. The crystals are washed with water and dried in vacuo: wt. 114 mg, M.P. 273-282". These crystals are designated C1. Chilling of I the mother liquor produces an additional 74 mg. of crystals 200 mg. of triamcinolone 16,17-acetophenonide for the triamcinolone 16,17-acetonide- 21-acetate,.the same product,'triamcinolone, is obtained.

which melt at ZZZ-244 and are designated C-2.

. Thin layer chromatography of C1 reveals that the bulk .of this material is unchanged triamcinolone acetonide.

This is corroborated by quantitative paper chromatography which shows that C-l contained 11% triamcinolone, 63 triamcinolone acetonide, and 30% of two faster moving spots consisting probably of triamcinolone 16,21-di- Z formats and triamcinolone acetonide 2l-formate respectively; C-2 consisted of 70% of-triamcinolone, 16% triamcinolone acetonide, and 14% triamcinolone diformate.

When a portion of C-1' (50 mg) is hydrolyzed with 10% potassium carbonate in methanol, crystallization 'of;

the resulting material yields'pure triamcinolone acetonide.

' EXAMPLE-13 A solution 013 .357 mg. of triamcinolone acetonide in 1? ml. of 88% formic acid is heatedon the steam cone at 100 for 30 minutes. After the remoyal of the reagent in vacuo followed by thorough drying for 1 hourat room temperature in vacuo, the residue is dissolved in 35 ml.

of methanol and under a blanket of nitrogen, 7 ml.'of 10% oxygen-free carbonate is added, the reaction mixture stirred for 1 hour under nitrogen and acidifiedwith 0.8 ml.

wherein the 1,2- and 6,7-positions are saturated or doublebonded, Ris hydrogen, R is selectedfrom the group consisting of hydrogen, hydroxy, and a-a'cyloxy, and together R and R is keto; each X is selected from the group con-;

sisting of hydrogen, halogen, hydroiry, lower alkoXy and lower allcyl, at least one X being selected from the group consisting of hydrogen and methyl; Y is selected from the group consisting ofhydrogen and methyl; Y isselected from the group consisting of hydrogen,. halogen and methyl; .Z is selected from the group consisting of by..-

droxyand acyloxygP is selected'from the group consisting of lower alkyl, halo-'loweralk'yl, carboXy-lower alkyl,

- monocyclic ,cycloalkyl, monocyclic aryljmonocyclic aryllower' alkyl, rnonocyclic heterocyclic and. monocyclic of'glacial acetic acid. ;After the addition of'water, the

solution is concentrated in Vacuo until crystallization 'occurs. Upon standing inthejrefrigerator, 162 mg. of material melting at 267270 called Cl is obtained. This. I materialis analyzed by quantitative'paper chromatography and is shown to consist of 98.5% pure triamcinolone, V

the remaining 1.5% being unchanged triamcinolone acetonide.

This invention may be variously otherwise embodied Within the'scope of the appended claims.

} Whatis claimed is: a

1. A process for preparing a l6et,i.7ot,2l-trihydroxy f steroid of the pregnaneseries which comprises treating a compound selectedfrom the group consisting of 16,17-'

cyclic ketals and acetals'of 16e,17a,2l-trihydroxy steroids of the pregnanev series and 2l-esters thereof .with an aqueous solution of formic acid containing more'than about 80% formic acid, hydrolyzing the formic acid ester formed, and recovering the 1604, 17a,21-trihydroxy steroid '1 formed.

. 2. The process of claim 1 wherein the steroid is of the.

general formulaheterocyclic lower alkyl; Q is selected from the group cons istingof hydrogen, loweralkyl; halo-lower alkyl,

carboxy-lower alkyl, monocyclic cycloalkyl, monocyclic aryl, monocyclic aryl lower alkyl, monocyclic heterocyclic and monocyclic heterocyclic lower alkyl; and together with the carbon'to which they are joined P and Q new I lected from the group consisting of cycloalkyl and monocyclic heterocyclic.

3. The. process of'clairn 1 wherein: the steroid is triamcinolone16,17-acetonide.

4. The processof claim -1 wherein the steroidisitri- A amcinolone- 16,17-acetonide 21- (lower alkanoate) 5. The process of claim 1 wherein thesteroid is triamcinolone16,17-acetonide ZI-acetate;

6. The process of'cl'aim :1 wherein the steroid is 6&-

flnoro-triamcinolone 16,17-acetonide 21,-(lower alkanoate). i i 7 7. They process ,of claim lwherein the steroid is 6w fluoro-triamcinolone l6, l7-acetonide .ZI-acetate;

8. The process of'claim 1 whereinthe; steroid iiSJ6ot- 'fltioro-16vt-hydroxyprednisolone 16,17-acetonide 21-(lowcr alkanoate).

3,021,341 2/62 Allen et 1, 260 297.45 3,050,519 8/62 Fried 260'239.55

. OTHER REFERENCES -Mills et al.: J.A.C.S. 81, pp. '126465(1959).'

LEWIS GOTTS, PrimaryjExaminer. 7 

1. A PROCESS FOR PREPARING A 16A,17A,21-TRIHYDROXY STEROID OF THE PREGNANE SERIES WHICH COMPRISES TREATING A COMPOUND SELECTED FROM THE GROUP CONSISTING OF 16,17CYCLIC KETALS AND ACETALS OF 16A,17A,21-TRIHYDROXY STEROIDS OF THE PREGNANE SERIES AND 21-ESTERS THEREOF WITH AN AQUEOUS SOLUTION OF FORMIC ACID CONTAINING MORE THAN ABOUT 80% FORMIC ACID, HYDROLYZING THE FORMIC ACID ESTER FORMED, AND RECOVERING THE 16A, 17A,21-TRIHYDROXY STERIOD FORMED. 